Department of Anatomical Sciences, Tarbiat Modares University, School of Medical Sciences, Tehran, Iran.
Restor Neurol Neurosci. 2010;28(3):367-77. doi: 10.3233/RNN-2010-0539.
Bone marrow stromal cells (BMSCs) are multipotent and can be induced to differentiate into different lineages including osteogenic, chondrogenic, myogenic and neuronal phenotypes. BMSCs were reported to be a suitable option for regenerative medical research. In this study, BMSCs were induced into GABAergic phenotypes using beta-mercaptoethanol (betaME) and retinoic acid (RA) as preinducers followed by potassium chloride as inducer, and were evaluated by fibronectin and Oct-4. The percentages of cells immunoreactive for nestin, neurofilaments (NF 68, NF 160, and NF 200), GABA and GABA synthesizing and packaging enzymes (GAD1, GAD2, VGAT) were used for evaluating GABAergic differentiation. RT-PCR was used for confirming the expression of these enzymes. The differentiated cells were loaded and unloaded with FM1-43 in order to assess the functionality of the cells. At the preinduction stage, the cells downregulated fibronectin and Oct-4, and expressed neuronal markers. At the induction stage, the preinduced cells transdifferentiated into GABAergic cells, as confirmed by immunohistochemistry and RT-PCR. The GABAergic cells were stained and destained with FM1-43. Therefore, the two-stage induction protocol resulted in transdifferentiation of BMSCs into GABAergic cells with synaptic release upon stimulation.
骨髓基质细胞（BMSCs）具有多能性，可以被诱导分化为不同的谱系，包括成骨细胞、软骨细胞、肌细胞和神经元表型。BMSCs 被报道是再生医学研究的一个合适选择。在这项研究中，BMSCs 先用β-巯基乙醇（βME）和维甲酸（RA）作为预诱导剂，再用氯化钾作为诱导剂，诱导分化为 GABA 能表型，并用纤维连接蛋白和 Oct-4 进行评估。巢蛋白、神经丝（NF68、NF160 和 NF200）、GABA 和 GABA 合成和包装酶（GAD1、GAD2、VGAT）的免疫反应性细胞的百分比用于评估 GABA 能分化。RT-PCR 用于确认这些酶的表达。分化的细胞被加载和卸载 FM1-43，以评估细胞的功能。在预诱导阶段，细胞下调纤维连接蛋白和 Oct-4，并表达神经元标志物。在诱导阶段，预诱导的细胞转分化为 GABA 能细胞，免疫组织化学和 RT-PCR 证实了这一点。GABA 能细胞用 FM1-43 染色和脱色。因此，两阶段诱导方案导致 BMSCs 转分化为 GABA 能细胞，并在刺激时释放突触。
